Imaging active site chemistry and protonation states: NMR crystallography of the tryptophan synthase α-aminoacrylate intermediate-Reference-Cited by-同舟云学术

Imaging active site chemistry and protonation states: NMR crystallography of the tryptophan synthase α-aminoacrylate intermediate

Published:2022-01-07 Issue:2 Volume:119 Page:
ISSN:0027-8424
Container-title:Proceedings of the National Academy of Sciences
language:en
Short-container-title:Proc. Natl. Acad. Sci. U.S.A.

Author:

Holmes Jacob B.¹,Liu Viktoriia¹,Caulkins Bethany G.¹²,Hilario Eduardo¹,Ghosh Rittik K.³,Drago Victoria N.⁴^ORCID,Young Robert P.⁵^ORCID,Romero Jennifer A.¹^ORCID,Gill Adam D.³,Bogie Paul M.¹,Paulino Joana⁶,Wang Xiaoling⁶^ORCID,Riviere Gwladys⁷^ORCID,Bosken Yuliana K.³^ORCID,Struppe Jochem⁸^ORCID,Hassan Alia⁹,Guidoulianov Jevgeni⁹,Perrone Barbara⁹,Mentink-Vigier Frederic⁶^ORCID,Chang Chia-en A.¹^ORCID,Long Joanna R.⁷,Hooley Richard J.¹³,Mueser Timothy C.⁴^ORCID,Dunn Michael F.³,Mueller Leonard J.¹^ORCID

Affiliation:

1. Department of Chemistry, University of California, Riverside, CA 92521;

2. W.M. Keck Science Department, Claremont McKenna, Pitzer, and Scripps Colleges, Claremont, CA 91711;

3. Department of Biochemistry, University of California, Riverside, CA 92521;

4. Department of Chemistry and Biochemistry, University of Toledo, Toledo, OH 43606;

5. Environmental Molecular Sciences Laboratory, Pacific Northwest National Laboratory, Richland, WA 99354;

6. National High Magnetic Field Laboratory, Florida State University, Tallahassee, FL 32310;

7. Department of Biochemistry and Molecular Biology, McKnight Brain Institute, National High Magnetic Field Laboratory, University of Florida, Gainesville, FL 32610;

8. Bruker Biospin Corporation, Billerica, MA 01821;

9. Bruker Switzerland AG 8117 Fällanden, Switzerland

Abstract

Significance The determination of active site protonation states is critical for a full mechanistic understanding of enzymatic transformations. However, hydrogen atom positions are challenging to extract using the standard tools of structural biology. Here, we make use of a joint solid-state NMR, X-ray crystallography, and first-principles computational approach that enables the investigation of enzyme catalysis at this fine level of chemical detail. For tryptophan synthase, this allows us to peer along the reaction coordinates into and out of the α-aminoacrylate intermediate. Through this process, we are developing a high-resolution probe for structural biology that is keenly sensitive to hydrogen atom positions—complementing diffraction methods yet able to be applied under conditions of active catalysis in microcrystalline and non-crystalline materials.

Funder

National Science Foundation

HHS | National Institutes of Health

Publisher

Proceedings of the National Academy of Sciences

Subject

Multidisciplinary

Link

https://pnas.org/doi/pdf/10.1073/pnas.2109235119

Reference84 articles.

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